Antibody drug conjugate purification

Abstract

The present invention relates to an ion exchange separation material with amino-acid based endgroups. This material is especially suitable for the separation and purification of ADCs.

1 . A separation material comprising a hydroxyl-group containing base matrix, to the surfaces of which polymer chains are grafted by covalent bonding, characterized in that the polymer chains comprise amino acid end groups —N(Y)—R3 with
 a. Y being independently from each other H or CH3, preferably H, and 
 b. R3 being —CHCOOMR4
 with R4 being C1 to C4 alkyl or C1 to C4 perfluoroalkyl 
 and M being H, Na, K, or NH 4 . 
 
 
     
     
         2 . Separation material according to  claim 1  whereby the monomer units of the polymer chains are linked in a linear manner and each monomer unit comprises an end group —N(Y)—R3. 
     
     
         3 . Separation material according to  claim 1  whereby the ionic density is between 400-900 μeq/g. 
     
     
         4 . Separation material according to  claim 1  whereby Y is H and R4 is isopropyl and/or isobutyl. 
     
     
         5 . Separation material according to  claim 1  whereby the hydroxylgroup containing base matrix comprises aliphatic hydroxyl groups. 
     
     
         6 . Separation material according to  claim 1  whereby the base matrix is formed by copolymerisation of a hydrophilically substituted alkyl vinyl ether selected from the group of 1,4-butanediol monovinyl ether, 1,5-pentanediol monovinyl ether, diethylene glycol monovinyl ether or cyclo¬hexane¬dimethanol monovinyl ether and divinylethyleneurea (1,3-divinylimidazolin-2-one) as crosslinking agent. 
     
     
         7 . Separation material according to  claim 1  whereby the separation material is preparable by subjecting the hydroxyl group containing base matrix to a cerium (IV) catalyzed graft polymerization of monomers according to formula V 
       
         
           
           
               
               
           
         
         with R1, R2 and Y being independently from each other H or CH3 
         R3 being —CHCOOMR4 
         with R4 being C1 to C4 alkyl or C1 to C4 perfluoroalkyl 
         and M being H, Na, K, or NH 4 . 
       
     
     
         8 . A process for the chromatographic purification and/or separation of antibody drug conjugates (ADCs) by contacting the sample comprising the ADCs with a separation material comprising a hydroxyl group containing base matrix, to the surfaces of which polymer chains are grafted by covalent bonding, characterized in that the polymers comprise end groups —N(Y)—R3 with
 Y being independently from each other H or CH 3    
 R3 being —CHCOOMR4 
 with R4 being C1 to C4 alkyl or C1 to C4 perfluoroalkyl, 
 and M being H, Na, K, or NH 4 . 
 
     
     
         9 . Process according to  claim 8  whereby the ADC target molecules are bound to the separation matrix at a pH between 2 and 7, optionally washed and eluted by pH change to alkaline pH. 
     
     
         10 . Process according to  claim 8  whereby the sample is applied to the separation matrix at an ionic density between 10-1200 μeq/g. 
     
     
         11 . Process according to  claim 8 , whereby between 10 mg and 100 mg ADCs are bound per ml of the separation material. 
     
     
         12 . Process for the preparation of a separation material according to  claim 1  by subjecting the hydroxyl group containing base matrix to a cerium (IV) catalyzed graft polymerization of monomers according to formula V 
       
         
           
           
               
               
           
         
         with R1, R2 and Y being independently from each other H or CH3, 
         R3 being —CHCOOMR4 
         with R4 being C1 to C4 alkyl or C1 to C4 perfluoroalkyl, and M being H, Na, K, or NH 4 .